Effect of water temperature on frog virus 3 disease in hatchery-reared pallid sturgeon Scaphirhynchus albus.

Ranaviruses are large double-stranded DNA viruses within the genus Ranavirus (family Iridoviridae) that are being detected with increasing frequency among aquacultured and wild fishes. In the USA, multiple sturgeon hatcheries have experienced ranavirus epizootics resulting in significant morbidity and mortality in young-of-year (YOY). Significant economic losses have resulted from repeated outbreaks of frog virus 3 (FV3), the type species for the genus Ranavirus, in YOY pallid sturgeon Scaphirhynchus albus reared at a hatchery within the Missouri River Basin. Water temperature and stocking density are known to influence the severity of ranavirus disease in ectothermic vertebrates. To determine the effect of water temperature on ranavirus disease in hatchery-raised S. albus, we conducted FV3 challenges at 2 temperatures (17 and 23°C) and compared cumulative survival over a 28 d study period. A mean (±SE) survival rate of 57.5 ± 13.2% was observed in replicate tanks of sturgeon maintained at 23°C, whereas no mortality was observed among sturgeon maintained at 17°C. In a second challenge study, we compared the effect of water temperature on disease progression by regularly sampling fish over the study period and evaluating lesions by histopathology and in situ hybridization, and by assessing viral titer and load in external and internal tissues using virus isolation and qPCR, respectively. Results suggest that temperature manipulation may be an effective mitigation strategy that sturgeon hatcheries can employ to minimize ranavirus-associated disease.

Multinucleate parietal cells and cytoplasmic inclusion bodies in the gastric epithelium of callitrichids.

Inclusion bodies (IBs) and multinucleate cells can be associated with viral infections; however, IBs and multinucleate cells have been described in normal tissue and with non-viral disease processes in multiple species. We examined fundic stomach from 50 callitrichids histologically for bi- and multinucleate parietal cells and cytoplasmic IBs in gastric epithelial cells. Callitrichids represented included 6 genera: Saguinus (4 spp.), Leontopithecus (1 sp.), Mico (3 spp.), Cebuella (1 sp.), Callithrix (1 sp.), Callimico (1 sp.), and 13 unspecified marmosets. Gastric epithelial IBs were present in 46 of 47 (98%) of the callitrichids from which the stomach was sufficiently well preserved to identify IBs. Cytoplasmic IBs were identified in gastric surface pit epithelial cells (43 of 44, 98%), mucous neck cells (43 of 44, 98%), parietal cells (43 of 44, 98%), and chief cells (43 of 44, 98%). The IBs were eosinophilic, ovoid, round, elongate, or variably indented, sometimes slightly refractile, and 1-6 × 1-13 µm. IBs were sometimes perinuclear and molded around the nucleus. Electron microscopy of the gastric epithelium of one marmoset indicated that IBs were composed of intermediate filaments. The IBs did not stain with immunohistochemical markers for cytokeratin AE1/AE3 or vimentin. Binucleate parietal cells were found in 49 of 50 (98%) callitrichids, and multinucleate parietal cells were observed in 40 of 49 (82%) callitrichids. Gastric epithelial cytoplasmic IBs and bi- and multinucleate parietal cells are likely a normal finding in callitrichids, and, to our knowledge, have not been reported previously.

POSTMORTEM COMPUTED TOMOGRAPHY AND MAGNETIC RESONANCE IMAGING FINDINGS IN A CASE OF COINFECTION OF DOLPHIN MORBILLIVIRUS AND ASPERGILLUS FUMIGATUS IN A JUVENILE BOTTLENOSE DOLPHIN (TURSIOPS TRUNCATUS).

A freshly dead juvenile bottlenose dolphin (Tursiops truncatus), recovered from the waters near Sand Key, Clearwater, FL, was imaged postmortem using computed tomography and magnetic resonance imaging prior to conventional necropsy. The pattern of imaging findings in the brain was compatible with severe multifocal meningoencephalitis with intralesional necrosis and/or hemorrhage, and the pattern of imaging findings in the lungs was compatible with severe multifocal bronchopneumonia. The subsequent investigation included necropsy, histology, culture, and molecular diagnostics and demonstrated disseminated coinfection of dolphin morbillivirus and Aspergillus fumigatus. This is the first report documenting the cross-sectional imaging findings of this important cetacean comorbidity and demonstrates advances in modern, cooperative investigations of marine mammal mortality events.

RENAL NEOPLASIA FROM A SINGLE POPULATION OF PTEROPODID BATS.

Neoplastic diseases have rarely been reported in the family Pteropodidae, and primary malignant renal neoplasms are generally uncommon across animal species. This case series describes four cases of primary renal neoplasia: three renal cell carcinomas and one nephroblastoma in three species of pteropodid bats, specifically large flying foxes (Pteropus vampyrus, n = 2), straw-colored fruit bat (Eidolon helvum, n = 1), and a little golden-mantled flying fox (Pteropus pumilus, n = 1). Two of the cases were diagnosed antemortem using ultrasonography, computed tomography, and cytology; and one of these bats with a renal cell carcinoma was treated successfully with a unilateral nephrectomy. The remaining two cases were diagnosed at necropsy.

Characterization of mule deerpox virus in Florida white-tailed deer fawns expands the known host and geographic range of this emerging pathogen.

Infections caused by mule deerpox virus (MDPV) have been sporadically reported in North American cervids. White-tailed deer (Odocoileus virginianus) fawns from a farm located in South Central Florida presented with ulcerative and crusting lesions on the coronary band as well as the mucocutaneous tissues of the head. Evaluation of the crusted skin lesions was undertaken using microscopic pathology and molecular techniques. A crusted skin sample was processed for virus isolation in four mammalian cell lines. The resulting isolate was characterized by negative staining electron microscopy and deep sequencing. Histopathologic evaluation of the skin lesions from the fawns revealed a hyperplastic and proliferative epidermis with ballooning degeneration of epidermal and follicular keratinocytes with intracytoplasmic eosinophilic inclusions. Electron microscopy of cell culture supernatant demonstrated numerous large brick-shaped particles typical of most poxviruses. Polymerase chain reaction assays followed by Sanger sequencing revealed a poxvirus gene sequence nearly identical to that of previous strains of MDPV. The full genome was recovered by deep sequencing and genetic analyses supported the Florida white-tailed deer isolate (MDPV-F) as a strain of MDPV. Herein, we report the first genome sequence of MDPV from a farmed white-tailed deer fawn in the South Central Florida, expanding the number of locations and geographic range in which MDPV has been identified.

EXTENSION OF THE KNOWN HOST RANGE OF INTRANUCLEAR COCCIDIOSIS: INFECTION IN THREE CAPTIVE RED-FOOTED TORTOISES ( CHELONOIDIS CARBONARIA).

The intranuclear coccidian parasite of Testudines (TINC) is an emerging pathogen of tortoises. Three captive red-footed tortoises ( Chelonoidis carbonaria) from an isolated collection presented with multiple acute, nonspecific clinical signs. One tortoise died and was diagnosed with intranuclear coccidiosis on histopathology with confirmation by quantitative polymerase chain reaction (qPCR). In addition to tissues where TINC has been previously described, coccidia were identified in the pineal gland, choroid plexus, and testicular Sertoli cells. The two remaining tortoises survived after treatment with oral ponazuril (20 mg/kg every 48 hr for 56 days) and remained asymptomatic, although not cleared of infection, for 21 months, as the number of coccidian gene copies detected by qPCR was reduced in one tortoise. This report extends the known host range of this parasite to continental South American tortoises, describes new sites of infection by histopathology, and has management implications for this disease.

Raccoons (Procyon lotor) as Sentinels of Trace Element Contamination and Physiological Effects of Exposure to Coal Fly Ash.

Anthropogenic pollutants disrupt global biodiversity, and terrestrial sentinels of pollution can provide a warning system for ecosystem-wide contamination. This study sought to assess whether raccoons (Procyon lotor) are sentinels of local exposure to trace element contaminants at a coal fly ash site and whether exposure resulted in health impairment or changes in the intestinal helminth communities. We compared trace element accumulation and the impact on health responses and intestinal helminth communities of raccoons inhabiting contaminated and reference sites of the U.S. Department of Energy's Savannah River Site (South Carolina, USA). Data on morphometry, hematology, histopathology, helminth community and abundance, and liver trace element burdens were collected from 15 raccoons captured adjacent to a coal fly ash basin and 11 raccoons from a comparable uncontaminated site nearby. Of eight trace elements analyzed, Cu, As, Se, and Pb were elevated in raccoons from the contaminated site. Raccoons from the contaminated site harbored higher helminth abundance than animals from the reference site and that abundance was positively associated with increased Cu concentrations. While we found changes in hematology associated with increased Se exposure, we did not find physiological or histological changes associated with higher levels of contaminants. Our results suggest that raccoons and their intestinal helminths act as sentinels of trace elements in the environment associated with coal fly ash contamination.

A retrospective study of the prevalence of calcium oxalate crystals in veterinary Aspergillus cases.

Fungi in the genus Aspergillus are some of the most common fungal pathogens in veterinary species, primarily affecting the respiratory tract. In both human and veterinary cases, calcium oxalate crystals have been documented in sites of Aspergillus infection. Cases in multiple species (16 birds, 15 horses, 5 dogs, 1 ox, and 1 dolphin) were identified that had either positive cultures for Aspergillus sp., or had conidiophores present that could be identified as belonging to the genus Aspergillus. Histologic slides were examined to confirm the presence of oxalate crystals and how often they were identified on the original report. Calcium oxalate deposition was detected in 14 of 38 cases examined, including A. fumigatus, A. versicolor, A. niger, and unspecified Aspergillus sp. infections. Calcium oxalate crystals were identified in 11 of 16 avian cases, as well as in 1 of 1 bovine, 1 of 15 equine, and 1 of 5 canine cases. Crystals were described in only 3 of the 14 original pathology reports of these cases, indicating that identification and reporting of crystals in histologic specimens could be improved. All the tissues with crystals were respiratory tissues with air interfaces, including nasal sinus, trachea, syrinx, lung, and air sac. In cases with crystals identified on H&E-stained sections, crystals were frequently not present or were fewer in number in tissue sections stained with Gomori methenamine silver and periodic acid-Schiff. Routine polarization of slides of fungal infections, especially in the respiratory tract, should be considered to check for calcium oxalate crystals.

Clinical behavior of intraocular teratoid medulloepithelioma in two-related Quarter Horses.

The objective of this paper is to describe clinical behavior, histopathologic features, and immunohistochemical staining of two-related horses with intraocular teratoid medulloepithelioma. Two-related Quarter Horses with similar intraocular masses presented to the UF-CVM Comparative Ophthalmology Service for evaluation and treatment. The first horse, a 3-year-old gelding, had glaucoma and a cyst-like mass in the anterior chamber. Enucleation was performed. Histopathology revealed a teratoid medulloepithelioma. The tumor was considered to be completely excised. Fifteen months later, the gelding presented with swelling of the enucleated orbit and local lymph nodes with deformation of the skull. Cytology revealed neuroectodermal neoplastic cells. Necropsy confirmed tumor metastasis. Six weeks later, a 9-year-old mare, a full sibling to the gelding, presented for examination. An infiltrative mass of the iris and ciliary body was found that extended into the anterior, posterior, and vitreal chambers. Uveitis was present, but secondary glaucoma was not noted. Enucleation was performed and the histopathologic diagnosis was also teratoid medulloepithelioma. The mare has had no recurrence to date, 2 years following enucleation. Metastasis of intraocular teratoid medulloepithelioma is possible. Staging is recommended in cases where the diagnosis of teratoid medulloepithelioma is confirmed. Surveillance of full siblings is recommended until more information regarding etiology is known.

Coinfection by Cetacean morbillivirus and Aspergillus fumigatus in a juvenile bottlenose dolphin (Tursiops truncatus) in the Gulf of Mexico.

A recently deceased juvenile male bottlenose dolphin (Tursiops truncatus) was found floating in the Gulf of Mexico, off Sand Key in Clearwater, Florida. At autopsy, we identified pneumonia and a focus of malacia in the right cerebrum. Cytologic evaluation of tissue imprints from the right cerebrum revealed fungal hyphae. Fungal cultures of the lung and brain yielded Aspergillus fumigatus, which was confirmed by amplification of a portion of the fungal nuclear ribosomal internal transcribed spacer 2 region sequence. Microscopic pulmonary lesions of bronchiolar epithelial cell syncytia with intracytoplasmic and intranuclear inclusions within bronchiolar epithelial cells were suggestive of Cetacean morbillivirus (CeMV) infection. The occurrence of CeMV infection was supported by positive immunohistochemical staining for morbillivirus antigen. CeMV detection was confirmed by amplification and sequencing a portion of the morbilliviral RNA-dependent RNA polymerase gene from lung tissue. This case provides CeMV sequence data available from the Gulf of Mexico and underscores the need for genomic sequencing across diverse host, temporospatial, and population (i.e., single animal vs. mass mortality events) scales to improve our understanding of these globally emerging pathogens.

A RETROSPECTIVE STUDY OF THE LESIONS ASSOCIATED WITH IRON STORAGE DISEASE IN CAPTIVE EGYPTIAN FRUIT BATS (ROUSETTUS AEGYPTIACUS).

Egyptian fruit bats (Rousettus aegyptiacus) are one of many species within zoologic collections that frequently develop iron storage disease. The goals of this retrospective multi-institutional study were to determine the tissue distribution of iron storage in captive adult Egyptian fruit bats and the incidence of intercurrent neoplasia and infection, which may be directly or indirectly related to iron overload. Tissue sections from 83 adult Egyptian fruit bats were histologically evaluated by using tissue sections stained with hematoxylin and eosin, trichrome, and Prussian blue techniques. The liver and spleen consistently had the largest amount of iron, but significant amounts of iron were also detected in the pancreas, kidney, skeletal muscle, and lung. Hepatocellular carcinoma (HCC; 11) was the most common neoplasm, followed by cholangiocarcinoma (4). Extrahepatic neoplasms included bronchioloalveolar adenoma (3), pulmonary carcinosarcoma (1), oral sarcoma (1), renal adenocarcinoma (1), transitional cell carcinoma of the urinary bladder (1), mammary gland adenoma (1), and parathyroid adenoma (1). There were also metastatic neoplasms of undetermined primary origin that included three poorly differentiated carcinomas, a poorly differentiated sarcoma, and a neuroendocrine tumor. Bats with hemochromatosis were significantly more likely to have HCC than bats with hemosiderosis (P = 0.032). Cardiomyopathy was identified in 35/77 bats with evaluable heart tissue, but no direct association was found between cardiac damage and the amount of iron observed within the liver or heart. Hepatic abscesses occurred in multiple bats, although a significant association was not observed between hemochromatosis and bacterial infection. To the authors' knowledge, this is the first publication providing evidence of a positive correlation between hemochromatosis and HCC in any species other than humans.

Evaluation of surgical outcome, complications, and mortality in dogs undergoing preoperative computed tomography angiography for diagnosis of an extrahepatic portosystemic shunt: 124 cases (2005-2014).

This study evaluated the safety of preoperative computed tomography angiography (CTA) and its effect on surgical time and clinical outcomes in dogs that underwent surgical correction of a single congenital extrahepatic portosystemic shunt (CEPSS). Patient data were retrospectively collected from medical records and owner communications for 124 dogs with single CEPSS, undergoing preoperative CTA (n = 43) or not (n = 81) which were surgically treated from 2005 to 2014. The frequency of major postoperative complications was 4.7% and 9.9% for the CTA and no CTA groups, respectively (P = 0.49). Mean ± standard deviation (SD) surgical time for the preoperative CTA group was 84 ± 40 min and 81 ± 31 min for the no CTA group (P = 0.28). We conclude that anesthetized preoperative CTA appears to be a safe method for diagnosis and surgical planning in dogs with single CEPSS, and does not appear to affect surgical procedure time, complication rate, or clinical outcome.

Évaluation des résultats chirurgicaux, des complications et de la mortalité chez les chiens subissant une angiographie par tomodensitométrie préopératoire pour le diagnostic d'un shunt portosystémique extrahépatique : 124 cas (2005­2014). Cette étude a évalué l'innocuité d'une angiographie par tomodensitométrie (AT) préopératoire et son effet sur la durée de la chirurgie et les résultats cliniques chez les chiens qui avaient subi la correction chirurgicale d'un shunt portosystémique extrahépatique congénital simple (SPSEHC). Les données des patients ont été recueillies rétrospectivement dans les dossiers médicaux et lors de communications avec les propriétaires pour 124 chiens atteints d'un SPSEHC simple, qui subissaient une AT préopératoire (n = 43) ou non (n = 81), et qui avaient été traités par chirurgie entre 2005 et 2014. La fréquence des complications postopératoires majeures était de 4,7 % et de 9,9 % pour les groupes AT et sans AT, respectivement (P = 0,49). La durée moyenne ± SD de la chirurgie pour le groupe d'AT préopératoire était de 84 ± 40 minutes et de 81 ± 31 minutes pour le groupe sans AT (P = 0,28). Nous avons conclu que l'AT préopératoire semble être une méthode sûre pour le diagnostic et la planification chirurgicale des chiens ayant un SPSEHC simple et qu'elle ne semble pas affecter la durée de l'intervention, le taux de complication ou les résultats cliniques.(Traduit par Isabelle Vallières).

RENAL AND CLOACAL CRYPTOSPORIDIOSIS (CRYPTOSPORIDIUM AVIAN GENOTYPE V) IN A MAJOR MITCHELL'S COCKATOO (LOPHOCHROA LEADBEATERI).

A 7-yr-old male Major Mitchell's cockatoo (Lophochroa leadbeateri) presented with a recent history of lethargy and anorexia. Physical examination revealed poor body condition and cloacal prolapse. Abnormalities on serum chemistry included severe hyperuricemia and hyperphosphatemia with a low calcium-to-phosphorus ratio. Symptomatic treatment was initiated including intravenous fluids and antibiotics. The bird continued to decline and died within a few days. Visceral gout and renal and cloacal pathology were observed on gross necropsy. Histopathology revealed chronic inflammation within the kidney, ureter, and cloaca in association with protozoal organisms and an invasive cloacal adenocarcinoma tumor. The location and morphology was consistent with Cryptosporidium sp., confirmed by immunohistochemistry and molecular testing. Direct sequencing identified Cryptosporidium avian genotype V. To the author's knowledge, this is the first reported infection of Cryptosporidium avian genotype V associated with clinical disease in birds and the first renal Cryptosporidium infection in a psittacine.

Bacterial sepsis resulting in severe systemic illness and euthanasia in a dog with cutaneous angiomatosis.

A Labrador retriever dog was euthanized after unsuccessful treatment for severe, progressive, lethargy, gastroenteritis, icterus, and swelling of a previously diagnosed cutaneous angiomatosis lesion. The body was submitted for necropsy. This is the first report that suggests that cutaneous angiomatosis lesions may have caused life-threatening systemic complications in a dog.

Septicémie bactérienne produisant une maladie systémique grave et l'euthanasie chez un chien atteint d'angiomatose cutanée. Un chien Labrador retriever a été euthanasié après l'échec du traitement pour un abattement sévère progressif, une gastro-entérite, l'ictère et l'enflure d'une lésion d'angiomatose cutanée diagnostiquée antérieurement. Le corps a été soumis à l'autopsie. C'est le premier rapport qui suggère que les lésions d'angiomatose cutanée peuvent avoir causé des complications systémiques potentiellement mortelles chez un chien.(Traduit par Isabelle Vallières).

The onset of goiter in several species of sharks following the addition of ozone to a touch pool.

This report describes the onset of goiter in several species of shark following the addition of ozone to a touch pool. A detailed description of a female brown-banded bamboo shark (Chiloscyllium punctatum) that was presented with multinodular goiter is provided. Four other brown-banded bamboo sharks and 11 white-spotted bamboo sharks (Chiloscyllium plagiosum) housed in the same system developed clinical disease consistent with goiter, but two zebra bullhead sharks (Heterodontus zebra) did not. Plasma thyroxine (T4) concentration was 4.64 ng/ml before euthanasia, consistent with a diagnosis of hypothyroidism. The sharks had been chronically exposed to mean (+/- standard error) NO3-N concentrations of 35 +/- 5.12 mg/L before ozonation of the system. Ozonation of aquarium water causes a reduction in environmental iodide, which is required for thyroid hormone synthesis. Nitrate is goitrogenic and would further decrease I- absorption by competitive inhibition. Multinodular goiter is consistent with goiter caused by chronic iodide deficiency. Understanding the interaction between water chemistry and goiter development is critical to development of elasmobranch health management systems.

Medium-grade astrocytoma in a cougar (Puma concolor).

A 17-year-old, male castrated cougar (Puma concolor) was presented minimally responsive and severely depressed, with bilateral mydriasis and absent pupillary light response. On gross examination of the brain, there was a tan-to-gray, invasive mass with a central cavitation on the ventral aspect in the left cerebral hemisphere, rostral to the caudate nucleus. On histopathologic examination, the mass was composed of sheets of medium-sized, round-to-polygonal cells that were multifocally separated by islands of neuropil. Approximately 80% of the neoplastic cells showed strong cytoplasmic labeling for glial fibrillary acidic protein. These findings were consistent with a medium-grade astrocytoma. To the authors' knowledge, neoplastic disease of the central nervous system has not been previously reported in cougars.

Incomplete ovariosalpingectomy and subsequent malignant granulosa cell tumor in a female green iguana (Iguana iguana).

CASE DESCRIPTION: A 9-year-old spayed female green iguana (Iguana iguana) was evaluated because of a distended coelom and weight loss. History included a single episode of egg binding and subsequent bilateral ovariosalpingectomy. CLINICAL FINDINGS: Physical examination revealed a mass within the coelomic cavity. Ultrasonography revealed a large, irregular mass with hypoechoic regions and coelomic effusion. Clinicopathologic derangements included heterophilia, monocytosis, lymphopenia, basophilia, hypocholesterolemia, hypoproteinemia, and hypercalcemia. Results of cytologic evaluation of the mass were suggestive of malignant epithelial neoplasia, but neoplastic cells were not found in the effusion. An ovarian tumor was suspected on the basis of clinical signs, clinicopathologic findings, and results of cytologic evaluation of the mass. TREATMENT AND OUTCOME: Surgical exploration revealed a large left ovary, a normal-appearing contralateral ovary, and a mass in the fat body, all of which were removed and submitted for histologic examination. The histologic diagnosis was granulosa cell tumor with metastasis to the fat body. The patient died 11 months after evaluation, and disseminated granulosa cell tumor was confirmed at necropsy; histologic examination at that time also identified systemic mastocytosis. CLINICAL RELEVANCE: Granulosa cell tumors are uncommon in reptiles, and this was the first granulosa cell tumor described antemortem cytologically, histologically, and ultrastructurally in an iguana. Findings in this iguana underscored concerns associated with incomplete oophorectomy of iguanas; cytologic and histopathologic findings were similar to those observed in other domestic animals. Oophorectomy should be considered as an alternative to standard ovariosalpingectomy to avoid potential complications in pet reptiles, and use of microsurgical instruments and vascular clips is advised.

Unilateral orbital lacrimal gland abscess in a horse.

A 20-year-old Thoroughbred gelding presented for evaluation of a periorbital dorsal swelling of the left eye that had been intermittently present for 3 months. Upon ocular examination, a firm, non-painful swelling was identified under the upper eyelid in the region of the orbital lacrimal gland, and was noted to extend anteriorly from underneath the dorsal orbital rim. Ultrasonographic examination revealed a mixed echogenic mass along the dorsal orbital rim that followed the contour of the globe. CT scan showed a moderately contrast enhancing mass that was contiguous with the eyelid. Differential diagnoses included neoplasia, inflammatory lesions such as a granuloma, foreign body or abscess. Surgical exploration and excision of the mass revealed a lobular structure with a purulent center. Histopathology identified the mass as the orbital lacrimal gland with concurrent severe dacryoadenitis. Culture of the purulent center of the mass revealed beta-hemolytic Staphylococcus aureus. The patient was maintained on supportive care and antibiotic treatment based on sensitivity postoperatively. No recurrence was reported 40 months later. This paper aims to identify bacterial dacryoadenitis as a cause for unilateral periorbital swelling in the horse. Differential diagnoses for this presentation, as well as successful surgical management are discussed. To the author's knowledge, this is the first case of bacterial dacryoadenitis and subsequent abscessation of the orbital lacrimal gland in the horse.